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Genetics Ch 6DNAREP

Genetics Ch 6DNAREP

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Published by April Iman
Genetic overview of DNA replication: from PSU Introduction to Genetics with prof. Estes
Genetic overview of DNA replication: from PSU Introduction to Genetics with prof. Estes

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Published by: April Iman on Oct 25, 2010
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Genetics Ch 6: Dna Replication + Modification25/10/2010 00:52:00
Linear DNA = multiple replication sitesMany OriRep – about 40,000 bases apart
Replicated in 15-30 minutesReplication of full genome (many chromosomes) = 5-10 hoursOkazaki fragments: 1000-2000bp in Prok, 100-200bp in Eukaryotic
Important Replication Proteins
Gyrase – topoisomerase II (double-stranded break)
Helicase – Unwinds DNA at replication fork to separate parentalstrands
SSB – stabilizes single-stranded DNA
Primosome – creates RNA primer
RNA Primase
DNA Polymerase – 5’->3’ Polymerization + proofreading functionsIn Euk,
RNA primer removed by RPA
(Replication Protein A)In Prok, RNA primer removal/replacement by DNA Pol I.
Both use DNA ligaseCircular DNA can replicate unidirectionally (rolling circle)(plasmids) orbidirectionally (theta)Lagging strand synth. leaves overhang once primer removed – telomeraseneeded. Telomerase contains built in RNA primer segment (reversetranscriptase activity)In
, only required enzyme is DNA Polymerase (primers are added)
Use special high-temperature DNA Pol from bacterial thermophileTAC Polymerase
Nucleid Acid Hybridization
: Denature DNA -> can combine with othercomplimentary or almost-complimentary strand. 
Restriction Enzymes
: Cut duplex DNA at particular sequences(endonucleases)
Often used as genome defense – antivirals. Host DNA is modified(methylation of C and A), unmodified DNA is vulnerable torestriction
Can be symmetric or asymmetric – useful because it leaves
sticky ends
Particular RE will chop a genome into a consistent set of 
restriction fragments
. Different RE = different
Chromosomal Basis of Heredity
Chp 3,2Females = homogametic (XX)Males = heterogametic (XY)
Punnett Square
Birds and butterflies = OPPOSITE, Males = ZZ, Females = ZWGregor Mendel – general rules of heredity.
1856-1863: pea experiments
First statistical rules
Elements of heredity (factors) remain unchanged thru generations
blending inheritance
Garden pea: lots of alternative traits, self-fertilization, easy toartificially fertilize
Lots of “selfing” = True/pure breeding = P generation
Monohybrid Cross
: cross bw types differing by one trait
Round vs Wrinkled
Determined by one gene
Encodes starch-branching enzyme 1 (
Synthesizes amylopectic
Round (W) seeds contain amylopectic – uniform shrinking
Wrinkled (w) seeds shrink irregularly (inborn error of starchmetabolism)
In w (wrinkled): mutation in transposable element interrupts gene
W smaller than w (transposable element,) can be separatedin a gel
Homologous genes are “wildtype” Blocking chlorophyll breakdown = peas stay green longer (yellow aredominant)After F1 hybrid self fertilization:
Green x Yellow = 3 yellow, 1 green (inheritance not sex-linked)YY
F2 ratios always like thisEx: Widow’s Peak: often in 3:1 ratio in humans. “Particulate” nature of heredity – gamete are equally likel to contain eachhalf of gene
Self cross
of F2:
¾ were round
1/3 give only round, 2/3 gave round and wrinkledin 3:1 ratio
¼ were wrinkled
all wrinkled
Test Cross
: Unknown genotype crossed w/ homozygous recessive “tester” 
Reveals genotype of unknown – if ½ wrinkled, ½ smooth unknown= Ww x ww = 2Ww, 2 wwDihybrid Cross: cross b/w types differing in 2 characters

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